Am. J. Respir. Cell Mol. Biol., Volume 22, Number 3, March, 2000 360-366

Induction and Regulation of Xenobiotic-Metabolizing Cytochrome P450s in the Human A549 Lung Adenocarcinoma Cell Line

Janne Hukkanen, Arja Lassila, Katja Päivärinta, Susanna Valanne, Sari Sarpo, Jukka Hakkola, Olavi Pelkonen, and Hannu Raunio

Department of Pharmacology and Toxicology, University of Oulu, Oulu, Finland

Several cytochrome P450 (CYP) enzymes are expressed in the human lung, where they participate in metabolic inactivation and activation of numerous exogenous and endogenous compounds. In this study, the expression pattern of all known xenobiotic-metabolizing CYP genes was characterized in the human alveolar type II cell-derived A549 adenocarcinoma cell line using qualitative reverse transcriptase/polymerase chain reaction (RT-PCR). In addition, the mechanisms of induction by chemicals of members in the CYP1 and CYP3A subfamilies were assessed by quantitative RT-PCR. The expression of messenger RNAs (mRNAs) of CYPs 1A1, 1B1, 2B6, 2C, 2E1, 3A5, and 3A7 was detected in the A549 cells. The amounts of mRNAs of CYPs 1A2, 2A6, 2A7, 2A13, 2F1, 3A4, and 4B1 were below the limit of detection. 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) induced CYP1A1 and CYP1B1 mRNAs 56-fold and 2.5-fold, respectively. CYP3A5 was induced 8-fold by dexamethasone and 11-fold by phenobarbital. CYP3A4 was not induced by any of the typical CYP3A4 inducers used. The tyrosine kinase inhibitor genistein and the protein kinase C inhibitor staurosporine blocked TCDD-elicited induction of CYP1A1, but they did not affect CYP1B1 induction. Protein phosphatase inhibitors okadaic acid and calyculin A enhanced TCDD-induction of CYP1B1 slightly, but had negligible effects on CYP1A1 induction. These results suggest that CYP1A1 and CYP1B1 are differentially regulated in human pulmonary epithelial cells and give the first indication of the induction of CYP3A5 by glucocorticoids in human lung cells. These results establish that having retained several characteristics of human lung epithelial cell CYP expression, the A549 lung cell line is a valuable model for mechanistic studies on induction of the pulmonary CYP system.

This article has been cited by other articles:

				J. S. Biggs, J. Wan, N. S. Cutler, J. Hakkola, P. Uusimaki, H. Raunio, and G. S. Yost Transcription Factor Binding to a Putative Double E-Box Motif Represses CYP3A4 Expression in Human Lung Cells Mol. Pharmacol., September 1, 2007; 72(3): 514 - 525. [Abstract] [Full Text] [PDF]

				N. Amara, R. Bachoual, M. Desmard, S. Golda, C. Guichard, S. Lanone, M. Aubier, E. Ogier-Denis, and J. Boczkowski Diesel exhaust particles induce matrix metalloprotease-1 in human lung epithelial cells via a NADP(H) oxidase/NOX4 redox-dependent mechanism Am J Physiol Lung Cell Mol Physiol, July 1, 2007; 293(1): L170 - L181. [Abstract] [Full Text] [PDF]

				N. Li and A. E. Nel The cellular impacts of diesel exhaust particles: beyond inflammation and death. Eur. Respir. J., April 1, 2006; 27(4): 667 - 668. [Full Text] [PDF]

				S. D. Spivack, G. J. Hurteau, M. J. Fasco, and L. S. Kaminsky Phase I and II Carcinogen Metabolism Gene Expression in Human Lung Tissue and Tumors Clin. Cancer Res., December 1, 2003; 9(16): 6002 - 6011. [Abstract] [Full Text] [PDF]

				J Millard, R B Parsons, R H Waring, A C Williams, and D B Ramsden Expression of cysteine dioxygenase (EC 1.13.11.20) and sulfite oxidase in the human lung: a potential role for sulfate production in the protection from airborne xenobiotica Mol. Pathol., October 1, 2003; 56(5): 270 - 274. [Abstract] [Full Text] [PDF]

				J. Hukkanen, T. Vaisanen, A. Lassila, R. Piipari, S. Anttila, O. Pelkonen, H. Raunio, and J. Hakkola Regulation of CYP3A5 by Glucocorticoids and Cigarette Smoke in Human Lung-Derived Cells J. Pharmacol. Exp. Ther., February 1, 2003; 304(2): 745 - 752. [Abstract] [Full Text] [PDF]

				J. M. Martinez, C. A. Afshari, P. R. Bushel, A. Masuda, T. Takahashi, and N. J. Walker Differential Toxicogenomic Responses to 2,3,7,8-Tetrachlorodibenzo-p-dioxin in Malignant and Nonmalignant Human Airway Epithelial Cells Toxicol. Sci., October 1, 2002; 69(2): 409 - 423. [Abstract] [Full Text] [PDF]

				N. T. Palackal, S. H. Lee, R. G. Harvey, I. A. Blair, and T. M. Penning Activation of Polycyclic Aromatic Hydrocarbon trans-Dihydrodiol Proximate Carcinogens by Human Aldo-keto Reductase (AKR1C) Enzymes and Their Functional Overexpression in Human Lung Carcinoma (A549) Cells J. Biol. Chem., June 28, 2002; 277(27): 24799 - 24808. [Abstract] [Full Text] [PDF]

				T. R. Van Vleet, K. Mace, and R. A. Coulombe Jr. Comparative Aflatoxin B1 Activation and Cytotoxicity in Human Bronchial Cells Expressing Cytochromes P450 1A2 and 3A4 Cancer Res., January 1, 2002; 62(1): 105 - 112. [Abstract] [Full Text] [PDF]

				T. R. Van Vleet, D. W. Bombick, and R. A. Coulombe Jr. Inhibition of Human Cytochrome P450 2E1 by Nicotine, Cotinine, and Aqueous Cigarette Tar Extract in Vitro Toxicol. Sci., December 1, 2001; 64(2): 185 - 191. [Abstract] [Full Text] [PDF]

				J. Hukkanen, O. Pelkonen, and H. Raunio Expression of xenobiotic-metabolizing enzymes in human pulmonary tissue: possible role in susceptibility for ILD Eur. Respir. J., July 1, 2001; 18(32_suppl): 122S - 126s. [Abstract] [Full Text] [PDF]

				M. J. Ronis, J. C. Rowlands, R. Hakkak, and T. M. Badger Inducibility of Hepatic CYP1A Enzymes by 3-Methylcholanthrene and Isosafrole Differs in Male Rats Fed Diets Containing Casein, Soy Protein Isolate or Whey from Conception to Adulthood J. Nutr., April 1, 2001; 131(4): 1180 - 1188. [Abstract] [Full Text]

				M. D. Roth, J. A. Marques-Magallanes, M. Yuan, W. Sun, D. P. Tashkin, and O. Hankinson Induction and Regulation of the Carcinogen-Metabolizing Enzyme CYP1A1 by Marijuana Smoke and Delta 9-Tetrahydrocannabinol Am. J. Respir. Cell Mol. Biol., March 1, 2001; 24(3): 339 - 344. [Abstract] [Full Text] [PDF]